Oligosaccharides and peptide displayed on an amphiphilic polymer enable solid phase assay of hapten specific antibodies.

Abstract

Copovidone, a copolymer of vinyl acetate and N-vinyl-2-pyrrolidone, was synthesized via reversible addition-fragmentation chain transfer (RAFT) polymerization, and after deacetylation the polymer was functionalized by introduction of amino, azide, and alkyne pendant groups to allow attachment of glycans and peptide. Candida albicans β-mannan trisaccharides 1 and 2 and M. tuberculosis arabinan hexasaccharide 3 with appropriate tethers were conjugated to the polymers by squarate or click chemistry. C. albicans T-cell peptide 4 bearing a C-terminal ε-azidolysine was also conjugated to copovidone by click chemistry. The resulting conjugates provide convenient non-protein-based antigens that are readily adsorbed on ELISA plates, and display excellent characteristics for assay of antibody binding to the haptenic group of interest. Copovidone and BSA glycoconjugates exhibited similar adsorption characteristics when used to coat ELISA plates, and both conjugates were optimal when used as coating solutions at low nanogram/mL concentrations. Provided that the copovidone conjugated glycan is stable to acid, assay plates can be easily processed for reuse at least three times without detectable variation or degradation in ELISA readout.

• Authors: David R Bundle, Pui-Hang Tam, Huu-Anh Tran, Eugenia Paszkiewicz, Jonathan Cartmell, Joanna M Sadowska, Susmita Sarkar, Maju Joe, Pavel I Kitov
• Journal: Bioconjugate chemistry (Bioconjug Chem) Vol. 25 Issue 4 Pg. 685-97 (Apr 16 2014) ISSN: 1520-4812 [Electronic] United States
• PMID: 24601638 (Publication Type: Journal Article, Research Support, Non-U.S. Gov't)

Chemical References

• Antibodies
• Haptens
• Oligosaccharides
• Peptides
• Polymers
• Pyrrolidines
• Surface-Active Agents
• Vinyl Compounds
• poly(vinylpyrrolidone-co-vinyl-acetate)

Topics

• Adsorption
• Antibodies (analysis, immunology)
• Antibody Specificity
• Candida albicans (chemistry, immunology)
• Click Chemistry
• Enzyme-Linked Immunosorbent Assay
• Equipment Reuse
• Haptens (chemistry, immunology)
• Molecular Conformation
• Mycobacterium tuberculosis (chemistry, immunology)
• Oligosaccharides (chemistry, immunology)
• Peptides (chemistry, immunology)
• Polymers (chemistry)
• Pyrrolidines (chemistry, immunology)
• Surface-Active Agents (chemistry)
• Vinyl Compounds (chemistry, immunology)